Home Hsa-miR-186-5p regulates TGFβ signaling pathway through expression suppression of SMAD6 and SMAD7 genes in colorectal cancer
Article
Licensed
Unlicensed Requires Authentication

Hsa-miR-186-5p regulates TGFβ signaling pathway through expression suppression of SMAD6 and SMAD7 genes in colorectal cancer

  • Zahra Bayat , Zahra Ghaemi , Mehrdad Behmanesh and Bahram M. Soltani ORCID logo
Published/Copyright: January 18, 2021
Biological Chemistry
From the journal Biological Chemistry Volume 402 Issue 4

Abstract

TGFβ signaling is a known pathway to be involved in colorectal cancer (CRC) progression and miRNAs play crucial roles by regulating different components of this pathway. Hence, finding the link between miRNAs and the pathway could be beneficial for CRC therapy. Array data indicated that miR-186-5p is a differentially expressed miRNA in colorectal Tumor/Normal tissues and bioinformatics tools predicted SMAD6/7 (inhibitory SMADs) as bona fide targets of this miRNA. Here, we intended to investigate the regulatory effect of the miR-186-5p expression on TGFβ signaling in CRC. Firstly, the miR-186-5p overexpression in HCT116 cells resulted in a significant reduction of SMAD6/7 expression, measured through RT-qPCR. Then, the direct interactions of miR-186-5p with SMAD6/7 3′UTRs were supported through dual luciferase assay. Furthermore, miR-186-5p overexpression suppressed proliferation, cell viability, and migration while, it increased apoptosis in CRC cells, assessed by cell cycle, MTT, scratch and Annexin V/PI apoptosis assays. Consistently, miR-186-5p overexpression resulted in reduced CyclinD1 protein using western blot, and also resulted in increased P21 and decreased c-Myc expression. Overall, these results introduced miR-186-5p as a cell cycle suppressor through downregulation of SMAD6/7 expression. Thus, miR-186-5p might be served as a novel tumor suppressive biomarker and therapeutic target in CRC treatment.

Keywords: colorectal cancer; miR-186; SMAD6; SMAD7; TGFβ signaling
Corresponding author: Bahram M. Soltani, Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran 14115-154, Islamic Republic of Iran, E-mail:
Zahra Bayat and Zahra Ghaemi contributed equally to this work.

Acknowledgments

Authors are thankful for the help and advices of all the members of 4402 laboratory in Genetics Departments at Tarbiat Modares University (Tehran, Iran).

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: None declared.

  3. Conflict of interest statement: The authors declare that they have no competing interests.

References

Agostini, M., Pucciarelli, S., Calore, F., Bedin, C., Enzo, M., and Nitti, D. (2010). miRNAs in colon and rectal cancer: a consensus for their true clinical value. Clin. Chim. Acta 411: 1181–1186. https://doi.org/10.1016/j.cca.2010.05.002.Search in Google Scholar PubMed

Butz, H., Rácz, K., Hunyady, L., and Patócs, A. (2012). Crosstalk between TGF-β signaling and the microRNA machinery. Trends Pharmacol. Sci. 33: 382–393. https://doi.org/10.1016/j.tips.2012.04.003.Search in Google Scholar PubMed

Cai, J., Wu, J., Zhang, H., Fang, L., Huang, Y., Yang, Y., Zhu, X., Li, R., and Li, M. (2013). miR-186 downregulation correlates with poor survival in lung adenocarcinoma, where it interferes with cell-cycle regulation. Canc. Res. 73: 756–766. https://doi.org/10.1158/0008-5472.can-12-2651.Search in Google Scholar PubMed

Cui, G., Cui, M., Li, Y., Liang, Y., Li, W., Guo, H., and Zhao, S. (2014). MiR-186 targets ROCK1 to suppress the growth and metastasis of NSCLC cells. Tumor Biol. 35: 8933–8937. https://doi.org/10.1007/s13277-014-2168-6.Search in Google Scholar PubMed

Derynck, R., Akhurst, R.J., and Balmain, A. (2001). TGF-β signaling in tumor suppression and cancer progression. Nat. Genet. 29: 117–129. https://doi.org/10.1038/ng1001-117.Search in Google Scholar PubMed

Di Guglielmo, G.M., Le Roy, C., Goodfellow, A.F., and Wrana, J.L. (2003). Distinct endocytic pathways regulate TGF-β receptor signalling and turnover. Nat. Cell Biol. 5: 410–421. https://doi.org/10.1038/ncb975.Search in Google Scholar PubMed

Erdmann, K., Kaulke, K., Thomae, C., Huebner, D., Sergon, M., Froehner, M., Wirth, M.P., and Fuessel, S. (2014). Elevated expression of prostate cancer-associated genes is linked to down-regulation of microRNAs. BMC Canc. 14: 82. https://doi.org/10.1186/1471-2407-14-82.Search in Google Scholar PubMed PubMed Central

Feng, H., Zhang, Z., Qing, X., French, S.W., and Liu, D. (2019). miR-186-5p promotes cell growth, migration and invasion of lung adenocarcinoma by targeting PTEN. Exp. Mol. Pathol. 108: 105–113. https://doi.org/10.1016/j.yexmp.2019.04.007.Search in Google Scholar PubMed

Genovese, G., Ergun, A., Shukla, S.A., Campos, B., Hanna, J., Ghosh, P., Quayle, S.N., Rai, K., Colla, S., Ying, H., et al.. (2012). microRNA regulatory network inference identifies miR-34a as a novel regulator of TGF-β signaling in glioblastoma. Canc. Discov. 2: 736–749. https://doi.org/10.1158/2159-8290.cd-12-0111.Search in Google Scholar

Geraldo, M.V., Yamashita, A.S., and Kimura, E.T. (2012). MicroRNA miR-146b-5p regulates signal transduction of TGF-β by repressing SMAD4 in thyroid cancer. Oncogene 31: 1910–1922. https://doi.org/10.1038/onc.2011.381.Search in Google Scholar PubMed

Gong, J., Ammanamanchi, S., Ko, T.C., and Brattain, M.G. (2003). Transforming growth factor β1 increases the stability of p21/WAF1/CIP1 protein and inhibits CDK2 kinase activity in human colon carcinoma FET cells. Canc. Res. 63: 3340–3346.Search in Google Scholar

Gu, S., Jin, L., Zhang, F., Sarnow, P., and Kay, M.A. (2009). Biological basis for restriction of microRNA targets to the 3′ untranslated region in mammalian mRNAs. Nat. Struct. Mol. Biol. 16: 144. https://doi.org/10.1038/nsmb.1552.Search in Google Scholar PubMed PubMed Central

Hayashi, H., Abdollah, S., Qiu, Y., Cai, J., Xu, Y.Y., Grinnell, B.W., Richardson, M.A., Topper, J.N., Gimbrone, M.A.Jr., Wrana, J.L., et al.. (1997). The MAD-related protein Smad7 associates with the TGFβ receptor and functions as an antagonist of TGFβ signaling. Cell 89: 1165–1173. https://doi.org/10.1016/s0092-8674(00)80303-7.Search in Google Scholar PubMed

Hlubek, F., Spaderna, S., Schmalhofer, O., Jung, A., Kirchner, T., and Brabletz, T. (2007). Wnt/FZD signaling and colorectal cancer morphogenesis. Front. Biosci. 12: 458–470. https://doi.org/10.2741/2075.Search in Google Scholar PubMed

Jafarzadeh, M., Soltani, B.M., Tousi, S.E., and Behmanesh, M. (2018). Hsa-miR-497 as a new regulator in TGFβ signaling pathway and cardiac differentiation process. Gene 675: 150–156. https://doi.org/10.1016/j.gene.2018.06.098.Search in Google Scholar PubMed

Jones, D.Z., Schmidt, M.L., Suman, S., Hobbing, K.R., Barve, S.S., Gobejishvili, L., Brock, G., Klinge, C.M., Rai, S.N., Park, J., et al.. (2018). Micro-RNA-186-5p inhibition attenuates proliferation, anchorage independent growth and invasion in metastatic prostate cancer cells. BMC Canc. 18: 421. https://doi.org/10.1186/s12885-018-4258-0.Search in Google Scholar PubMed PubMed Central

Katsuno, Y., Lamouille, S., and Derynck, R. (2013). TGF-β signaling and epithelial–mesenchymal transition in cancer progression. Curr. Opin. Oncol. 25: 76–84. https://doi.org/10.1097/cco.0b013e32835b6371.Search in Google Scholar

Ko, T.C., Sheng, H.M., Reisman, D., Thompson, E.A., and Beauchamp, R.D. (1995). Transforming growth factor-beta 1 inhibits cyclin D1 expression in intestinal epithelial cells. Oncogene 10: 177–184.Search in Google Scholar

Liu, J., Zheng, M., Tang, Y.L., Liang, X.H., and Yang, Q. (2011). MicroRNAs, an active and versatile group in cancers. Int. J. Oral Sci. 3: 165–175. https://doi.org/10.4248/ijos11063.Search in Google Scholar PubMed PubMed Central

Liu, Z., Zhang, G., Yu, W., Gao, N., and Peng, J. (2016). miR-186 inhibits cell proliferation in multiple myeloma by repressing Jagged1. Biochem. Biophys. Res. Commun. 469: 692–697. https://doi.org/10.1016/j.bbrc.2015.11.136.Search in Google Scholar PubMed

Markowitz, S.D. and Roberts, A.B. (1996). Tumor suppressor activity of the TGF-β pathway in human cancers. Cytokine Growth Factor Rev. 7: 93–102. https://doi.org/10.1016/1359-6101(96)00001-9.Search in Google Scholar PubMed

Massagué, J. (2000). How cells read TGF-β signals. Nat. Rev. Mol. Cell Biol. 1: 169–178. https://doi.org/10.1038/35043051.Search in Google Scholar PubMed

Massagué, J. (2008). TGFβ in cancer. Cell 134: 215–230. https://doi.org/10.1016/j.cell.2008.07.001.Search in Google Scholar PubMed PubMed Central

Massagué, J., Blain, S.W., and Lo, R.S. (2000). TGFβ signaling in growth control, cancer, and heritable disorders. Cell 103: 295–309. https://doi.org/10.1016/s0092-8674(00)00121-5.Search in Google Scholar PubMed

McGuire, S. (2016). World cancer report 2014. World Health Organization, International Agency for Research on Cancer, WHO Press, Geneva, Switzerland, 2015.10.3945/an.116.012211Search in Google Scholar

Miyazawa, K. and Miyazono, K. (2017). Regulation of TGF-β family signaling by inhibitory Smads. Cold Spring Harb. Perspect. Biol. 9: a022095. https://doi.org/10.1101/cshperspect.a022095.Search in Google Scholar PubMed PubMed Central

Moez, M.J., Bjeije, H., and Soltani, B.M. (2019). Hsa-miR-5195-3P induces downregulation of TGFβR1, TGFβR2, SMAD3 and SMAD4 supporting its tumor suppressive activity in HCT116 cells. Int. J. Biochem. Cell Biol. 109: 1–7. https://doi.org/10.1016/j.biocel.2019.01.001.Search in Google Scholar PubMed

Mukherjee, P., Winter, S.L., and Alexandrow, M.G. (2010). Cell cycle arrest by transforming growth factor β1 near G1/S is mediated by acute abrogation of prereplication complex activation involving an Rb-MCM interaction. Mol. Cell Biol. 30: 845–856. https://doi.org/10.1128/mcb.01152-09.Search in Google Scholar PubMed PubMed Central

Myatt, S.S., Wang, J., Monteiro, L.J., Christian, M., Ho, K.K., Fusi, L., Dina, R.E., Brosens, J.J., Ghaem-Maghami, S., and Lam, E.W. (2010). Definition of microRNAs that repress expression of the tumor suppressor gene FOXO1 in endometrial cancer. Canc. Res. 70: 367–377. https://doi.org/10.1158/0008-5472.can-09-1891.Search in Google Scholar PubMed PubMed Central

Nagaraj, N.S. and Datta, P.K. (2010). Targeting the transforming growth factor-β signaling pathway in human cancer. Expet Opin. Invest. Drugs 19: 77–91. https://doi.org/10.1517/13543780903382609.Search in Google Scholar PubMed PubMed Central

Pais, H., Nicolas, F.E., Soond, S.M., Swingler, T.E., Clark, I.M., Chantry, A., Moulton, V., and Dalmay, T. (2010). Analyzing mRNA expression identifies Smad3 as a microRNA-140 target regulated only at protein level. RNA 16: 489–494. https://doi.org/10.1261/rna.1701210.Search in Google Scholar PubMed PubMed Central

Shi, Y. and Massagué, J. (2003). Mechanisms of TGF-β signaling from cell membrane to the nucleus. Cell 113: 685–700. https://doi.org/10.1016/s0092-8674(03)00432-x.Search in Google Scholar PubMed

Shi, M., Zhu, J., Wang, R., Chen, X., Mi, L., Walz, T., and Springer, T.A. (2011). Latent TGF-β structure and activation. Nature 474: 343–349. https://doi.org/10.1038/nature10152.Search in Google Scholar PubMed PubMed Central

Siegel, P.M. and Massagué, J. (2003). Cytostatic and apoptotic actions of TGF-β in homeostasis and cancer. Nat. Rev. Canc. 3: 807–820. https://doi.org/10.1038/nrc1208.Search in Google Scholar PubMed

Slattery, M.L., Wolff, R.K., and Lundgreen, A. (2015). A pathway approach to evaluating the association between the CHIEF pathway and risk of colorectal cancer. Carcinogenesis 36: 49–59. https://doi.org/10.1093/carcin/bgu213.Search in Google Scholar PubMed PubMed Central

Xu, C., Li, B., Zhao, S., Jin, B., Jia, R., Ge, J., and Xu, H. (2019). MicroRNA-186-5p inhibits proliferation and metastasis of esophageal cancer by mediating HOXA9. OncoTargets Ther. 12: 8905. https://doi.org/10.2147/ott.s227920.Search in Google Scholar

Yagi, K., Furuhashi, M., Aoki, H., Goto, D., Kuwano, H., Sugamura, K., Miyazono, K., and Kato, M. (2002). c-myc is a downstream target of the Smad pathway. J. Biol. Chem. 277: 854–861. https://doi.org/10.1074/jbc.m104170200.Search in Google Scholar PubMed

Yao, K., He, L., Gan, Y., Zeng, Q., Dai, Y., and Tan, J. (2015). MiR-186 suppresses the growth and metastasis of bladder cancer by targeting NSBP1. Diagn. Pathol. 10: 1–7. https://doi.org/10.1186/s13000-015-0372-3.Search in Google Scholar PubMed PubMed Central

Ye, J.J. and Cao, J. (2014). MicroRNAs in colorectal cancer as markers and targets: recent advances. World J. Gastroenterol. 20: 4288. https://doi.org/10.3748/wjg.v20.i15.4288.Search in Google Scholar PubMed PubMed Central

Yu, J., Lei, R., Zhuang, X., Li, X., Li, G., Lev, S., Segura, M.F., Zhang, X., and Hu, G. (2016). MicroRNA-182 targets SMAD7 to potentiate TGFβ-induced epithelial-mesenchymal transition and metastasis of cancer cells. Nat. Commun. 7: 1–12. https://doi.org/10.1038/ncomms13884.Search in Google Scholar PubMed PubMed Central

Zhang, Y., Li, M., Wang, H., Fisher, W.E., Lin, P.H., Yao, Q., and Chen, C. (2009). Profiling of 95 microRNAs in pancreatic cancer cell lines and surgical specimens by real-time PCR analysis. World J. Surg. 33: 698. https://doi.org/10.1007/s00268-008-9833-0.Search in Google Scholar PubMed PubMed Central

Zhang, Z., Zhang, W., Mao, J., Xu, Z., and Fan, M. (2019). miR-186-5p functions as a tumor suppressor in human osteosarcoma by targeting FOXK1. Cell. Physiol. Biochem. 52: 553–564. https://doi.org/10.33594/000000039.Search in Google Scholar PubMed

Received: 2019-11-03
Accepted: 2020-12-28
Published Online: 2021-01-18
Published in Print: 2021-03-26

© 2021 Walter de Gruyter GmbH, Berlin/Boston

Articles in the same Issue

  1. Frontmatter
  2. Minireview
  3. Regulation of RNA stability at the 3′ end
  4. Research Articles/Short Communications
  5. Protein Structure and Function
  6. Phage-display reveals interaction of lipocalin allergen Can f 1 with a peptide resembling the antigen binding region of a human γδT-cell receptor
  7. Membranes, Lipids, Glycobiology
  8. Apolipoprotein C-II and C-III preferably transfer to both high-density lipoprotein (HDL)2 and the larger HDL3 from very low-density lipoprotein (VLDL)
  9. Molecular Medicine
  10. Dihydroartemisinin ameliorates balloon injury-induced neointimal formation through suppressing autophagy in vascular smooth muscle cells
  11. Cell Biology and Signaling
  12. HDAC8 promotes daunorubicin resistance of human acute myeloid leukemia cells via regulation of IL-6 and IL-8
  13. Hsa-miR-186-5p regulates TGFβ signaling pathway through expression suppression of SMAD6 and SMAD7 genes in colorectal cancer
  14. Lysine acetyltransferase Tip60 acetylates the APP adaptor Fe65 to increase its transcriptional activity
  15. Vorinostat exhibits anticancer effects in triple-negative breast cancer cells by preventing nitric oxide-driven histone deacetylation
  16. Human glucose-dependent insulinotropic polypeptide (GIP) is an antimicrobial adjuvant re-sensitising multidrug-resistant Gram-negative bacteria
  17. Retraction Note
  18. Retraction note to: LncRNA MEG3 inhibits HMEC-1 cells growth, migration and tube formation via sponging miR-147
https://doi.org/10.1515/hsz-2019-0407
Keywords for this article
colorectal cancer; miR-186; SMAD6; SMAD7; TGFβ signaling

Articles in the same Issue

  1. Frontmatter
  2. Minireview
  3. Regulation of RNA stability at the 3′ end
  4. Research Articles/Short Communications
  5. Protein Structure and Function
  6. Phage-display reveals interaction of lipocalin allergen Can f 1 with a peptide resembling the antigen binding region of a human γδT-cell receptor
  7. Membranes, Lipids, Glycobiology
  8. Apolipoprotein C-II and C-III preferably transfer to both high-density lipoprotein (HDL)2 and the larger HDL3 from very low-density lipoprotein (VLDL)
  9. Molecular Medicine
  10. Dihydroartemisinin ameliorates balloon injury-induced neointimal formation through suppressing autophagy in vascular smooth muscle cells
  11. Cell Biology and Signaling
  12. HDAC8 promotes daunorubicin resistance of human acute myeloid leukemia cells via regulation of IL-6 and IL-8
  13. Hsa-miR-186-5p regulates TGFβ signaling pathway through expression suppression of SMAD6 and SMAD7 genes in colorectal cancer
  14. Lysine acetyltransferase Tip60 acetylates the APP adaptor Fe65 to increase its transcriptional activity
  15. Vorinostat exhibits anticancer effects in triple-negative breast cancer cells by preventing nitric oxide-driven histone deacetylation
  16. Human glucose-dependent insulinotropic polypeptide (GIP) is an antimicrobial adjuvant re-sensitising multidrug-resistant Gram-negative bacteria
  17. Retraction Note
  18. Retraction note to: LncRNA MEG3 inhibits HMEC-1 cells growth, migration and tube formation via sponging miR-147
Sign up now to receive a 20% welcome discount
Institutional Access
How does access work?
Have an idea on how to improve our website?
Please write us.
© 2025 De Gruyter Brill
Downloaded on 9.9.2025 from https://www.degruyterbrill.com/document/doi/10.1515/hsz-2019-0407/html
Scroll to top button